Virus-Based Pyroelectricity.

The first observation of heat-induced electrical potential generation on a virus and its detection through pyroelectricity are presented. Specifically, the authors investigate the pyroelectric properties of the M13 phage, which possesses inherent dipole structures derived from the noncentrosymmetric arrangement of the major coat protein (pVIII) with an α-helical conformation. Unidirectional polarization of the phage is achieved through genetic engineering of the tail protein (pIII) and template-assisted self-assembly techniques. By modifying the pVIII proteins with varying numbers of glutamate residues, the structure-dependent tunable pyroelectric properties of the phage are explored. The most polarized phage exhibits a pyroelectric coefficient of 0.13 µC m-2 °C-1 . Computational modeling and circular dichroism (CD) spectroscopy analysis confirm that the unfolding of α-helices within the pVIII proteins leads to changes in phage polarization upon heating. Moreover, the phage is genetically modified to enable its pyroelectric function in diverse chemical environments. This phage-based approach not only provides valuable insights into bio-pyroelectricity but also opens up new opportunities for the detection of various viral particles. Furthermore, it holds great potential for the development of novel biomaterials for future applications in biosensors and bioelectric materials.

Elastic Fluorescent Protein-Based Down-Converting Optical Films for Flexible Display.

Protein-based material design provides great advantages to developing smart biomaterials with tunable structures and desired functions. They have been widely used in many biomedical applications including tissue engineering and drug delivery. However, protein-based materials are not yet widely used in optoelectronic materials despite their excellent optical and tunable mechanical properties. Here, we synthesized engineered fluorescent proteins (FPs) fused with elastic protein for the development of optoelectrical down-converting optical filters for flexible display materials. We synthesized sequence-specific FPs to tune blue, green, yellow, and red colors and fused them with elastic protein to tune mechanical properties. We fabricated flexible self-supporting film materials and characterized mechanical properties and down-converting optical properties. We also fabricated a hybrid light-emitting diode (LED) to down convert blue to desired green, red, and white colors. Furthermore, we constructed a flexible white LED using organic LED as a flexible substrate. Our modular synthesis approach of tunable bio-optoelectrical material approaches will be useful to design future biocompatible and flexible display materials and technologies.

Biomimetic virus-based soft niche for ischemic diseases.

The essential therapeutic cues provided by a nanofibrous arginine-glycine-aspartic acid-engineered M13 phage were exploited as extracellular matrix (ECM)-mimicking niches, contributing to de novo soft tissue niche engineering. The interplay of biomimetic phage cues with surrounding organ tissues was identified, and cells were implanted between tissues to achieve an appropriate soft tissue niche that enables the proper functioning of the implanted stem cells at the injured site. With the polyacrylamide (PA) hydrogel mimicking the soft tissue organ stiffness ranges, it was found that biochemical and topological cues in conjunction with the ∼1-2 kPa elastic and mechanical cues of engineered phage nanofibers in soft tissues efficiently enhance the desired response of implanted stem cells. This phage cue with angiogenic and antioxidant functions overcomes the pathological environment to support implanted cells and surrounding soft tissues at the ischemic site, thereby successfully decreasing myogenic degeneration, minimizing fibrosis, and enhancing blood vessel regeneration with M2 macrophage polarization by improving the survival of the implanted endothelial progenitor cells (EPC) in an ischemic mouse model. These biomimetic phage nanofiber cues are considerably supportive of cell therapy, as they establish promising therapeutic extracellular de novo soft tissue niches for curing ischemic diseases.

Recombinant factor VIII protein aggregation and adsorption at the liquid-solid interface.

Undesired aggregation and adsorption of therapeutic proteins during manufacturing and administration processes can significantly decrease the efficacy of protein drugs, especially when a quantitative treatment is critical. In this study, we investigate molecular interactions of recombinant factor VIII (rFVIII), a therapeutic protein for hemophilia A treatment, at a static liquid-glass interface. We quantitatively analyze the adsorption and aggregation of rFVIII using atomic force microscopy (AFM), dynamic light scattering (DLS) and UV-Vis spectroscopy. We also investigate how PEGylation, temperature, ionic strength and pH affect the rFVIII aggregation and adsorption at the interface over time. The aggregation and adsorption of rFVIII are significantly reduced by decreasing electrostatic attractions in the solution. We observed that the PEGylation endows rFVIII molecules with high stability at the liquid-glass interface in a wide range of temperature, ionic strength and pH. Our studies will help to understand the molecular interactions of how proteins aggregate and adsorb on the solid surface and prevent the undesired events in pharmaceutical applications.

Moisture-induced autonomous surface potential oscillations for energy harvesting.

A variety of autonomous oscillations in nature such as heartbeats and some biochemical reactions have been widely studied and utilized for applications in the fields of bioscience and engineering. Here, we report a unique phenomenon of moisture-induced electrical potential oscillations on polymers, poly([2-(methacryloyloxy)ethyl] dimethyl-(3-sulfopropyl) ammonium hydroxide-co-acrylic acid), during the diffusion of water molecules. Chemical reactions are modeled by kinetic simulations while system dynamic equations and the stability matrix are analyzed to show the chaotic nature of the system which oscillates with hidden attractors to induce the autonomous surface potential oscillation. Using moisture in the ambient environment as the activation source, this self-excited chemoelectrical reaction could have broad influences and usages in surface-reaction based devices and systems. As a proof-of-concept demonstration, an energy harvester is constructed and achieved the continuous energy production for more than 15,000 seconds with an energy density of 16.8 mJ/cm2. A 2-Volts output voltage has been produced to power a liquid crystal display toward practical applications with five energy harvesters connected in series.

M13 Virus Triboelectricity and Energy Harvesting.

Triboelectrification is a phenomenon that generates electric potential upon contact. Here, we report a viral particle capable of generating triboelectric potential. M13 bacteriophage is exploited to fabricate precisely defined chemical and physical structures. By genetically engineering the charged structures, we observe that more negatively charged phages can generate higher triboelectric potentials and can diffuse the electric charges faster than less negatively charged phages can. The computational results show that the glutamate-engineered phages lower the LUMO energy level so that they can easily accept electrons from other materials upon contact. A phage-based triboelectric nanogenerator is fabricated and it could produce ∼76 V and ∼5.1 µA, enough to power 30 light-emitting diodes upon a mechanical force application. Our biotechnological approach will be useful to understand the electrical behavior of biomaterials, harvest mechanical energy, and provide a novel modality to detect desired viruses in the future.

Catechol-Functionalized Elastin-like Polypeptides as Tissue Adhesives.

Adhesives can potentially be used to achieve fast and efficient wound closure; however, current products show poor bonding on wet surfaces, undergo swelling, and lack adequate biocompatibility. We designed a hydrogel adhesive with recombinant elastin-like polypeptides (ELPs), which are flexible proteins that can be customized for biomedical needs. The adhesive proteins are synthesized by chemically modifying the ELPs with dopamine, which contain adhesive catechol moieties. The resulting catechol-functional ELPs or Cat-ELPs can form flexible hydrogels that show stable swelling in aqueous conditions at 37 °C. We demonstrate their flexibility and viscoelastic properties through rheology. We also show the advantage of using customizable recombinant proteins to improve the material biological properties by demonstrating improved fibroblast binding on Cat-ELP by adding ELP with "RGD" peptides. We further confirmed in vivo biocompatibility and biodegradation of Cat-ELP hydrogels by implanting them in mice and monitoring for 10 weeks. Finally, we tested the bonding strength of the adhesives on porcine skin through tensile pull-off and lap-shear testing, in which we found strengths of 37 and 39 kPa, respectively. We demonstrated the tensile bonding strength by suspending a 2 kg mass on a one square inch (6.5 cm2) skin sample. As our adhesives are developed further, our strategy combining recombinant protein engineering and chemical modification will help yield an ideal bioadhesive for wound closure.

Effect of elastin-like polypeptide incorporation on the adhesion maturation of mineral trioxide aggregates.

The aim of this study was to investigate the effects of the incorporation of elastin-like polypeptide (ELP) on the adhesion maturation of mineral trioxide aggregates (MTA). Two types of ELPs (V125 and V125E8) were genetically synthesized. V125 consisted of 125 repeating pentapeptides (Val-Pro-Gly-Xaa-Gly) and V125E8 was functionalized with octaglutamic acid in the C-terminus of V125; both were diluted to 10 wt% in solution. Three 1.5 mm diameter holes in dentin discs were filled with MTA mixed with either a solution of ELP or deionized water. Push-out bond strength tests were performed following storage in simulated body fluid (SBF) for 1, 2, 4, and 8 weeks (n = 12). The interface between dentin and MTA was observed via scanning electron microscopy (SEM). The maturation of MTA was evaluated with a stereoscopic microscope and SEM. The incorporation of a specific ELP (V125E8) significantly increased the bond strength of MTA to dentin with regard to every maturation period (p < 0.05). The bond strength of MTA also significantly increased with a longer maturation time irrespective of ELP incorporation (p < 0.05). V125E8-incorporated MTA exhibited a more intimate interface with dentin compared to the other groups. More spindle-shaped crystal structures and thicker crystals were observed in all MTA mixtures as the storage duration increased even though V125E8 exhibited fewer crystal structures on the surface. Within the limitations of this study, the incorporation of V125E8 increased the adhesive properties of MTA and the maturation of MTA occurred regardless of ELP incorporation.

Biomolecular Piezoelectric Materials: From Amino Acids to Living Tissues.

Biomolecular piezoelectric materials are considered a strong candidate material for biomedical applications due to their robust piezoelectricity, biocompatibility, and low dielectric property. The electric field has been found to affect tissue development and regeneration, and the piezoelectric properties of biological materials in the human body are known to provide electric fields by pressure. Therefore, great attention has been paid to the understanding of piezoelectricity in biological tissues and its building blocks. The aim herein is to describe the principle of piezoelectricity in biological materials from the very basic building blocks (i.e., amino acids, peptides, proteins, etc.) to highly organized tissues (i.e., bones, skin, etc.). Research progress on the piezoelectricity within various biological materials is summarized, including amino acids, peptides, proteins, and tissues. The mechanisms and origin of piezoelectricity within various biological materials are also covered.

Elastin-Based Thermoresponsive Shape-Memory Hydrogels.

A shape-memory hydrogel is a programmable hydrogel material that can store specific shapes and execute functions in response to stimuli. In this report, we developed shape-memory hydrogels by creating double-network polymeric structures using a physically cross-linking elastin-like polypeptide (ELP) and a chemically cross-linking polyacrylamide (PAM). We synthesized the hydrogel matrix by polymerizing the acrylamide mixed in an ELP solution. We exploited the lower critical solution temperature transition of the ELP to enable the hydrogel to hold a new desired shape at an elevated temperature of 55 °C. The original shape of the hydrogel can then be recovered by lowering the temperature to 20 °C. The shape-memory hydrogels we developed exhibit ultrafast functionality and high repeatability. Taking advantage of the temperature-induced shape-memory capability, we also demonstrate practical functions such as gripping an object and connecting two tubes. Our materials with effective temperature-driven shape-memory functionality will be useful for developing novel materials for biomedical applications in the future.

Correction: Engineered phage nanofibers induce angiogenesis.

Correction for 'Engineered phage nanofibers induce angiogenesis' by So Young Yoo et al., Nanoscale, 2017, 9, 17109-17117.

Experimental and numerical evaluation of a genetically engineered M13 bacteriophage with high sensitivity and selectivity for 2,4,6-trinitrotoluene.

Selective and sensitive detection of desired targets is very critical in sensor design. Here, we report a genetically engineered M13 bacteriophage-based sensor system evaluated by quantum mechanics (QM) calculations. Phage display is a facile way to develop the desired peptide sequences, but the resulting sequences can be imperfect peptides for binding of target molecules. A TNT binding peptide (WHW) carrying phage was self-assembled to fabricate thin films and tested for the sensitive and selective surface plasmon resonance-based detection of TNT molecules at the 500 femtomole level. SPR studies performed with the WHW peptide and control peptides (WAW, WHA, AHW) were well-matched with those of the QM calculations. Our combined method between phage engineering and QM calculation will significantly enhance our ability to design selective and sensitive sensors.

Vertical Self-Assembly of Polarized Phage Nanostructure for Energy Harvesting.

Controlling the shape, geometry, density, and orientation of nanomaterials is critical to fabricate functional devices. However, there is limited control over the morphological and directional characteristics of presynthesized nanomaterials, which makes them unsuitable for developing devices for practical applications. Here, we address this challenge by demonstrating vertically aligned and polarized piezoelectric nanostructures from presynthesized biological piezoelectric nanofibers, M13 phage, with control over the orientation, polarization direction, microstructure morphology, and density using genetic engineering and template-assisted self-assembly process. The resulting vertically ordered structures exhibit strong unidirectional polarization with three times higher piezoelectric constant values than that of in-plane aligned structures, supported by second harmonic generation and piezoelectric force microscopy measurements. The resulting vertically self-assembled phage-based piezoelectric energy harvester (PEH) produces up to 2.8 V of potential, 120 nA of current, and 236 nW of power upon 17 N of force. In addition, five phage-based PEH integrated devices produce an output voltage of 12 V and an output current of 300 nA, simply by pressing with a finger. The resulting device can operate light-emitting diode backlights on a liquid crystal display. Our approach will be useful for assembling many other presynthesized nanomaterials into high-performance devices for various applications.

MoS2 Liquid Cell Electron Microscopy Through Clean and Fast Polymer-Free MoS2 Transfer.

Two dimensional (2D) materials have found various applications because of their unique physical properties. For example, graphene has been used as the electron transparent membrane for liquid cell transmission electron microscopy (TEM) due to its high mechanical strength and flexibility, single-atom thickness, chemical inertness, etc. Here, we report using 2D MoS2 as a functional substrate as well as the membrane window for liquid cell TEM, which is enabled by our facile and polymer-free MoS2 transfer process. This provides the opportunity to investigate the growth of Pt nanocrystals on MoS2 substrates, which elucidates the formation mechanisms of such heterostructured 2D materials. We find that Pt nanocrystals formed in MoS2 liquid cells have a strong tendency to align their crystal lattice with that of MoS2, suggesting a van der Waals epitaxial relationship. Importantly, we can study its impact on the kinetics of the nanocrystal formation. The development of MoS2 liquid cells will allow further study of various liquid phenomena on MoS2, and the polymer-free MoS2 transfer process will be implemented in a wide range of applications.

Bacteriophage nanofiber fabrication using near field electrospinning.

M13 bacteriophage (phage) nano- and microfibers were fabricated using electrospinning. Using liquid crystalline suspension of the phage, we successfully fabricated nano- and microscale pure phage fibers. Through a near field electrospinning process, we fabricated the desired phage fiber pattern with tunable direction and spacing. In addition, we demonstrated that the resulting phage fibers could be utilized as an electrostatic-stimulus responsive actuator. The near field electrospinning would be a very useful tool to design phage-based chemical sensors, tissue regenerative materials, energy generators, metallic and semiconductor nanowires in the future.

Enhancing Effect of Elastinlike Polypeptide-based Matrix on the Physical Properties of Mineral Trioxide Aggregate.

INTRODUCTION: Elastinlike polypeptide (ELP) is 1 of the genetically engineered, protein-based polypeptides, which offers outstanding advantages such as superior biocompatibility, long-term stability, elasticity, and cost-effectiveness. This study aimed to investigate the effect of an ELP-based matrix on the physical properties and biocompatibility of mineral trioxide aggregate (MTA). METHODS: The 2 types of ELPs were synthesized and mixed with the MTA powder in various liquid-to-powder ratios. The physical properties including compressive strength, microhardness and setting time, washout resistance, and biocompatibility were investigated for the ELP-incorporated MTA. The microstructure of the MTA was also analyzed using scanning electron microscopy and Fourier-transform infrared spectroscopy. RESULTS: The ELP-based matrix enhanced the physical properties of MTA, including the compressive strength, microhardness, and washout resistance of MTA. The ELP incorporation showed no negative effect on biocompatibility. However, ELPs prolonged the setting time of MTA. CONCLUSIONS: These results suggested that the addition of the ELP-based matrix to MTA enhanced the physical properties without negatively affecting the chemical structure and biocompatibility of MTA. Further investigation is warranted to overcome a clinical challenge associated with the extended setting time caused by the addition of ELP.

Diphenylalanine Peptide Nanotube Energy Harvesters.

Piezoelectric materials are excellent generators of clean energy, as they can harvest the ubiquitous vibrational and mechanical forces. We developed large-scale unidirectionally polarized, aligned diphenylalanine (FF) nanotubes and fabricated peptide-based piezoelectric energy harvesters. We first used the meniscus-driven self-assembly process to fabricate horizontally aligned FF nanotubes. The FF nanotubes exhibit piezoelectric properties as well as unidirectional polarization. In addition, the asymmetric shapes of the self-assembled FF nanotubes enable them to effectively translate external axial forces into shear deformation to generate electrical energy. The fabricated peptide-based piezoelectric energy harvesters can generate voltage, current, and power of up to 2.8 V, 37.4 nA, and 8.2 nW, respectively, with 42 N of force, and can power multiple liquid-crystal display panels. These peptide-based energy-harvesting materials will provide a compatible energy source for biomedical applications in the future.

Chimeric Adeno-Associated Virus-Mediated Cardiovascular Reprogramming for Ischemic Heart Disease.

Here, we demonstrated chimeric adeno-associated virus (chimeric AAV), AAV-DJ-mediated cardiovascular reprogramming strategy to generate new cardiomyocytes and limit collagen deposition in cardiac fibroblasts by inducing synergism of chimeric AAV-expressing Gata4, Mef2c, Tbx5 (AAV-GMT)-mediated heart reprogramming and chimeric AAV-expressing thymosin ß4 (AAV-Tß4)-mediated heart regeneration. AAV-GMT promoted a gradual increase in expression of cardiac-specific genes, including Actc1, Gja1, Myh6, Ryr2, and cTnT, with a gradual decrease in expression of a fibrosis-specific gene, procollagen type I and here AAV-Tß4 help to induce GMT expression, providing a chimeric AAV-mediated therapeutic cell reprogramming strategy for ischemic heart diseases.

Engineering of M13 Bacteriophage for Development of Tissue Engineering Materials.

M13 bacteriophages have several qualities that make them attractive candidates as building blocks for tissue regenerating scaffold materials. Through genetic engineering, a high density of functional peptides and proteins can be simultaneously displayed on the M13 bacteriophage's outer coat proteins. The resulting phage can self-assemble into nanofibrous network structures and can guide the tissue morphogenesis through proliferation, differentiation and apoptosis. In this manuscript, we will describe methods to develop major coat-engineered M13 phages as a basic building block and aligned tissue-like matrices to develop regenerative nanomaterials.

Growth of Au and ZnS nanostructures via engineered peptide and M13 bacteriophage templates.

We demonstrate directed nucleation of Au and ZnS patterns on templates comprised of functional peptides and an M13 bacteriophage. We discuss the control over nucleation in terms of the interplay between enhanced ion binding and reduced interfacial energy resulting from the presence of the templates.